放射线对体外培养棘球蚴及其Gadd45α mRNA表达的影响
Effects of radiation on echinococcus in vitro culture and its mRNA expression of Gadd45α
摘要目的:观察放射线对体外培养棘球蚴的杀伤作用及对其生长阻滞和DNA损伤45α(growth arrest and DNA damage 45 alpha,Gadd45α)mRNA表达的影响。方法:体外培养自然感染的羊肝脏中的棘球蚴,分为7组,分别以0(对照)、20、40、60、80、100、120 Gy的6兆电子线(6 MeV)进行1次放射。放射线照射后培养1、3、5、7 d时,光镜下观察棘球蚴的生长情况;采用反转录-聚合酶链反应(RT-PCR)法测定放射线照射后培养7 d时,对照,20、40、60 Gy组棘球蚴Gadd45α mRNA表达水平。结果:各剂量组放射线照射后1、3、5、7 d时,光镜下棘球蚴结构崩解,勾突脱落,且棘球蚴死亡率与放射线照射剂量呈正相关( r = 0.81, P < 0.05)。放射线照射后培养7 d时,与对照组(100.00 ± 0.00)比较,20、40、60 Gy组棘球蚴Gadd45α mRNA表达水平明显升高(279.74 ± 80.08、759.38 ± 160.98、1 666.68 ± 316.36, P均 < 0.01),且与放射线剂量呈正相关( r = 0.93, P < 0.01)。 结论:放射线照射对体外培养的棘球蚴有一定的杀伤作用,且与放射线的剂量存在一定的量效关系;Gadd45α基因可能参与放射线所致体外杀伤棘球蚴的分子机制。
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abstractsObjective:To investigate the killing effect of radiation on echinococcus in vitro culture and its effect on the mRNA expression of growth arrest and DNA damage 45 alpha (Gadd45α) gene. Methods:Echinococcus from naturally infected sheep liver was cultured in vitro and divided into 7 groups. The echinococcus was irradiated with 6 MeV at doses of 0 (control), 20, 40, 60, 80, 100, and 120 Gy, respectively. The growth of echinococcus was observed under light microscope at 1, 3, 5 and 7 d after the radiation. The expression of Gadd45α mRNA in control, 20, 40 and 60 Gy groups of echinococcus was detected by RT-PCR technique at 7 d after the radiation. Results:The disintegration and exfoliation of echinococcus were observed under light microscope at 1, 3, 5 and 7 d after the radiation, and the death rate of echinococcus was positively correlated with the radiation dosages ( r = 0.81, P < 0.05). After the radiation at 7 d, compared with the control group (100.00 ± 0.00), the mRNA expression levels of Gadd45α in echinococcus of 20, 40, and 60 Gy groups were significantly increased (279.74 ± 80.08, 759.38 ± 160.98, 1 666.68 ± 316.36, P < 0.01), and it was positively correlated with the radiation dosages ( r = 0.93, P < 0.01). Conclusion:Radiation has a certain killing effect on echinococcus cultured in vitro, and there is a certain dose-effect relationship with the radiation dosages, and Gadd45α gene may be involved in the molecular mechanism of radiation-induced killing of echinococcus in vitro.
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